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Koji mold, which belongs to the Aspergillus section Nigri, is used in the production of shochu. The section Nigri is composed of very morphologically similar members that in some cases produce mycotoxins, which rises concerns as to whether the presence of mycotoxin-producing fungi in shochu producing sites can compromise consumer safety. Thus, we examined the presence of mycotoxin-producing sec. Nigri fungi in six shochu factories (named A-F) in Japan. Airborne fungal levels in the factories were determined, and a traditional koji called "kona-koji" made from the mold naturally present in factory C (Aogashima village) was analyzed. Isolates of sec. Nigri fungi were identified morphologically and confirmed via cytochrome b gene analysis. In factory A (Nago city), airborne fungal levels of sec. Nigri were 4,000 and 100 cfu/m3 in the koji-making and fermentation rooms, respectively. In factories B, C, and D, the levels were 40, >104 cfu/m3, and 100 cfu/m3, respectively. In factory F (Iki city), there were high levels of airborne white-koji mold (a white mutant of Asp. luchuensis). The most dominant fungal species of sec. Nigri was isolated and identified as Asp. luchuensis via genetic analysis. This is likely to have originated from the commercial fermentation culture used. Asp. niger and Asp. luchuensis were isolated from kona-koji. Mycotoxin production (ochratoxin and fumonisin B2) by Asp. luchuensis (eight strains) and Asp. niger (three strains) was virtually inexistent; only one strain of Asp. niger was positive for fumonisin B2. This study clearly shows that mycotoxin-producing fungi are not dominant in the fungal flora present in the shochu factories examined and therefore, that the liquor can be safely fermented.Implications: In this study, we examined the presence of mycotoxin-producing Aspergillus sec. Nigri fungi in six shochu (Japanese distilled beverage) factories. The most dominant fungal species of sec. Nigri was isolated and identified as Aspergillus luchuensis (black-koji mold). The proportion of mycotoxin-producing Aspergillus niger and Aspergillus carbonarius was very small. In addition, the Asp. niger isolated from koji mold did not have the ability to produce ochratoxins or fumonisin B. This study clearly shows that shochu can be safely fermented.

Introduction. Dried fruits are a valuable source of dietary fibre, many vitamins and minerals in the population’s diet. However, the high content of readily available carbohydrates makes this type of product vulnerable to mould contamination. The greatest danger among which are toxigenic species. But there is practically no scientific information about the contamination presented on the Russian market dried fruits with moulds producing mycotoxins. That does not allow judging about this aspect of food safety. Materials and methods. Contamination with moulds and bacteria of 57 samples of dried fruits of 7 species popular in Russia was studied using cultural methods of analysis. Monospore isolates of moulds were isolated from dried fruits; in vitro mycotoxins production studied; by UHPLC-MS / MS analyzed mycotoxins in the multidetection mode. Results and discussion. In general, the microbial contamination of dried fruits was low: 87.7% of the samples met the established microbiological standards, in most cases, moulds caused it. At the same time, the highest frequency and levels of contamination were found in dates. Aspergillus sp. dominated in the micoflora of all types of dried fruits. Among the isolated 33 strains of moulds, 45.5% turned out to be toxigenic and, in vitro, were capable of biosynthesis of significant amounts of several types of mycotoxins, including emergent mycotoxins. Fumonisin- and ochratoxin-producing activities have been found in Aspergillus strains of the Nigri section. In model experiments, the accumulation of mycotoxins in individual strains exceeded the level normalized in grain products, including (in μg/kg): for aflatoxins B1 - more than 32000 and B2 - 3230; fumonisin B2 - more than 3100; ochratoxin A up to 4.3; for emergent accumulation reached: sterigmatocystin up to 6218220 and citreoviridine - 153. Conclusion. Moulds are the main type of microflora that contaminates dried fruits. The ability of mould isolates from dried fruits to form mycotoxins has been established, among which highly toxigenic strains have been identified. This indicates the presence of a potential risk of contamination of this type of food with unregulated mycotoxins and a possible increase in their content in the diets of consumers. The results obtained substantiate the need for extensive monitoring of mycotoxin producers in dried fruits. This is important for predicting the risk of toxin formation and identifying the relationship of specific mycotoxins with certain types of dried fruits. The presence of toxigenic activity of moulds isolated from dried fruits has been shown in Russia for the first time. Contribution: Minaeva L.P. — concept and design of the study, collection and processing of material of microbiological studies, writing and editing of the manuscript, responsibility for the integrity of all parts of the article; Polyanina A.S., Efimochkina N.R. — collection and processing of material of microbiological studies; Kiseleva M.G., Chalyy Z.A. — collection and processing of material UHPLC-MS/MS research results; Sheveleva S.A. — editing. All authors — approval of the final version of the article. Conflict of interest. The authors declare no conflict of interest. Acknowledgement. The study was supported by Russian Science Foundation (grant No 18‐16‐00077).

The aim of this study was to isolate Aspergillus section Nigri from onion samples bought in supermarkets and to analyze the fungal isolates by means of molecular data in order to differentiate A. niger and A. welwitschiae species from the other non-toxigenic species of black aspergilli, and detect genes involved in the biosynthesis of ochratoxin A and fumonisin B2. Aspergillus section Nigri were found in 98% (94/96) of the onion samples. Based on the results of multiplex PCR (performed on 500 randomly selected strains), 97.4% of the Aspergillus section Nigri strains were recognized as A. niger/A. welwitschiae. Around half of them were subjected to partial sequencing of the CaM gene to distinguish one from the other.Atotal of 97.9% of the isolates were identified as A. welwitschiae and only 2.1% as A. niger. The fum8 gene, involved in fumonisin B2 biosynthesis, was found in 36% of A. welwitschiae isolates, but radH and pks genes, involved in ochratoxin A biosynthesis, were found in only 2.8%. The presence/absence of fum8 gene in the A. welwitschiae genome is closely associated with ability/inability of the isolates to produce fumonisin in vitro. Based on these results,we suggest that in-depth studies are conducted to investigate the presence of fumonisins in onion bulbs. FAPESP / CAPES / CNPq

Nine domestic wine samples collected from a Japanese winery were examined for the presence of fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), as well as ochratoxin A (OTA) and ochratoxin B (OTB). Wine samples spiked with 13C-labeled internal standards (13C34-FB1 and 13C20-OTA) were diluted with phosphate buffered saline (PBS) buffer, loaded on immunoaffinity cartridges to purify of fumonisins and ochratoxins, and subjected to liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis. The data revealed that the domestic wine samples were possibly contaminated with FB1 and FB3, in addition to FB2, whereas none of the tested wine samples were contaminated with OTA and OTB. These results suggest that Fusarium fungi can be associated with the fumonisin contamination of Japanese domestic wine, whereas Aspergillus niger seems to be frequently reported as the major causal fungus of fumonisin contamination of wine in Europe. Analysis of the intermediate samples during the wine brewing indicated that fumonisin concentrations did not increase during wine production, suggesting that fumonisin contamination did not occur during the brewing process, but was derived from the raw materials of grape berries.

Aspergillus spp. are a common contaminant of grapes and a major source of mycotoxins. China is the largest producer of grape in the world now, however, the toxigenic Aspergillus population on grape in this country is still largely unknown. In this study, a total of 345 strains were isolated from grapes of 13 main grapevine producing regions in China. Based on calmodulin gene sequences, eight species within Aspergillus section Nigri were identified. Among them, Aspergillus tubingensis (48.7%) was predominant, followed by Aspergillus welwitschiae (20.6%) and Aspergillus aculeatinus (11%). Average of contamination level was up to 64.19%, and we found the occurrence of section Nigri species on the surface of fresh grapes was significantly influenced by the climate (P2 (FB2) producers. Of those, Aspergillus carbonarius was the main OTA producer and A. welwitschiae and Aspergillus niger were the main FB2 producers with different toxigenic ability. Our results highlight the potential risk of OTA and FB2 contamination by A. carbonarius and A. welwitschiae on grape in China, respectively; management strategies should be considered for preventing and reducing the toxigenic Aspergillus and its mycotoxins.

The garlic contains sulfur bioactive compounds responsible for medicinal properties. The decrease of these compounds due to inadequate storage conditions reduces the beneficial properties and favors infection by microorganisms. Several studies have shown high frequency of garlic infected with Aspergillus section Nigri that potentially produce mycotoxin. Garlic samples were collected in markets of Brazil and a total of 32 samples (of 36) had the fungal infection with predominant genus Aspergillus (50.3%), Penicillium (34.7%), and Fusarium (11%). A total of 63% (649/1031) of infection with Aspergillus section Nigri, of which 60 isolates were selected for analysis of genetic variability that resulted in 4 clusters. Representatives of clusters were identified by the calmodulin gene. Isolates from cluster I were subdivided into A-I and identified as A. niger (16 isolates) and the isolates of clusters B-I, II, and III were identified as A. welwitschiae (43 isolates). Besides, an isolate of the IV-cluster was identified by A. luchuensis. Further, we used the multiplex PCR to verify genotypes of 59 isolates, and none of these had OTA production-associated genotype. Moreover, 19 A. welwitschiae and 15 A. niger were FB2 production-associated genotype. Our study is the first report to the incidence of garlic infection in Brazil and to show that A. welwitschiae causes most of these infections.

The toxicokinetics of the food and feed contaminant Fumonisin B (FB) are characterized by low oral absorption and rapid plasma elimination. For these reasons, FB is not considered to accumulate in animals. However, recent studies in chicken and turkey showed that, in these species, the hepatic half-elimination time of fumonisin B1 (FB1) was several days, suggesting that FB1 may accumulate in the body. For the present study, 21-day-old chickens received a non-toxic dose of around 20 mg FB1 + FB2/kg of feed to investigate whether FB can accumulate in the body over time. Measurements taken after four and nine days of exposure revealed increased concentrations of sphinganine (Sa) and sphingosine (So) over time in the liver, but no sign of toxicity and no effect on performances were observed at this level of FB in feed. Measurements of FB in tissues showed that FB1 accumulated in chicken livers from four to nine days, with concentrations of 20.3 and 32.1 ng FB1/g observed, respectively, at these two exposure periods. Fumonisin B2 (FB2) also accumulated in the liver, from 0.79 ng/g at four days to 1.38 ng/g at nine days. Although the concentrations of FB found in the muscles was very low, an accumulation of FB1 over time was observed in this tissue, with concentrations of 0.036 and 0.072 ng FB1/g being measured after four and nine days of exposure, respectively. Feeding algo-clay to the chickens reduced the accumulation of FB1 in the liver and muscle by , approximately 40 and 50% on day nine, respectively. By contrast, only a weak non-significant effect was observed on day four. The decrease in the concentration of FB observed in tissues of chickens fed FB plus algo-clay on day nine was accompanied by a decrease in Sa and So contents in the liver compared to the levels of Sa and So measured in chickens fed FB alone. FB1 in the liver and Sa or So contents were correlated in liver tissue, confirming that both FB1 and Sa are suitable biomarkers of FB exposure in chickens. Further studies are necessary to determine whether FB can accumulate at higher levels in chicken tissues with an increase in the time of exposure and in the age of the animals.

The fungal diseases in the vineyard reduce grape quality influencing the volatile profile and consequently impair the aroma, taste, and color of juice and wine. In addition to this sensory effect, the occurrence of filamentous fungi in grapes may result in mycotoxins, including alternariol, aflatoxins, tenuazonic acid, fumonisin B2, ochratoxin A (OTA), patulin citrinin, among others. OTA is the most studied mycotoxin in grape products and can effectively represent a concern for industries. This short review discloses that promising results have been obtained with the use of antagonistic microorganisms or natural products for the toxigenic fungi control in the vineyard. Reduction of OTA levels throughout the production stages of juice and wine is well documented in the literature. However, studies taking into account the impacts of climate change on vine and the resilience of toxigenic fungi are still a challenge. In addition, the co-occurrence of OTA (including modified forms) and other mycotoxins in grapes products of different cultivars and regions needs to be evaluated to verify the risk of human exposure to these compounds.

This study was conducted to detect and quantify three mycotoxins Dioxynivalenol, Nivalenol and Fumonisin B2 in seeds of seven wheat cultivars planted in 17 wheat fields in Basra province, Iraq. This was done by using High Performance Liquid chromatographs analysis. The results revealed that Fumonisin B2 was the predominant mycotoxin, which present in 10 fields. The lowest concentration rate of this mycotoxin was 110 µg/Kg and the maximum was 11,228 µg/Kg. Dioxynivalenol as a trichothecene was in the second level detected in 6 fields with a minimum concentration of 8 µg/Kg and a maximum of 1,060 µg/Kg. Nivalenol was found only in 4 fields ranging from 272-1900 µg/Kg. Fumonisin B2 Only three fields showed co-occurrence of two mycotoxins (Fumonisin B2 and Nivalenol) in each, but with various concentration rates. The seven cultivars tested in this study were varied in their reactions to subjected mycotoxins. Adana 99 (A. 99) cultivar showed the highest concentration rate of both Fumonisin B2 and Nivalenol, which present with average percentage of 64% and 58% respectively. While, for Dioxynivalenol, Ebaa 99 (E.99) was on the top, it occurred in 6 fields ranging from 8-1060 µg/Kg with an average percentage of 43%.

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin, and rapid and sensitive detection of AFB1 is in demand for food safety and environmental analysis. Here we described a simple aptamer molecular beacon assay for rapid detection of aflatoxin B1 (AFB1) by using an aptamer with a fluorescein (FAM) label at the 5′ end and a fluorescence quencher (black hole quencher 1, BHQ1) at the 3′ end. In the presence of AFB1, the aptamer probe bound with AFB1 and induced a hairpin structure, drawing FAM and BHQ1 into close proximity and leading to fluorescence quenching. This assay allowed for a detection limit of 3.9 nmol/L and a dynamic range from 3.9 nmol/L to 4 μmol/L. Specificity test showed other mycotoxins including ochratoxin A, ochratoxin B, fumonisin B1, fumonisin B2, and zearalenone had negligible influence on detection of AFB1. AFB1 spiked in diluted liquor wine, methanol, or corn flour samples was successfully detected by using this aptamer probe, and the assay showed potential for real sample analysis.

Our study investigated the occurrence of a wide range of mycotoxins in coffee, most of them rarely analyzed in this commodity, and the safety relevance of mycotoxin exposure from coffee consumption. A new liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the analysis of 31 regulated and emerging mycotoxins was therefore developed and validated. Quantification was performed by the matrix-matched calibration curve approach. Analyses were conducted on green coffee which permitted to reach low limits of quantification, i.e. from 0.25 μg/kg to 150 μg/kg. A total of 71 green coffee beans samples, harvested in 2015 and 2016, were collected from various geographical origins (nine countries) and comprised of Arabica or Robusta species. Types of post-harvest processes and storage times in bags were also considered during sample collection. Diversity, occurrence and levels of mycotoxins in the surveyed green coffee samples were quite low. The mycotoxins most frequently found and at highest levels were Ochratoxin A, Fumonisin B2, Sterigmatocystin, Beauvericin and Enniatin A with maximum levels at 12.2, 76.3, 19.8, 4.4 and 1.7 μg/kg, respectively. Contamination with Aflatoxin B1 was sporadic with maximum level at 1.2 μg/kg. Co-occurrence of multiple mycotoxins per sample was rare, at very low levels and mostly limited to two different toxins. A risk assessment performed on our data set indicates that coffee is a minor contributor to mycotoxin exposure from the diet and is not expected to represent a safety concern.

Ülkemizde mısır üretimindeki artış bir yandan sürerken, bir yandan da dünyada mısırın soyadan sonra en çok genetiği değiştirilmiş ürün olması nedeniyle tüketicinin mısır ile ilgili algısı her geçen gün daha olumsuz olmaktadır. Genetiği değiştirilmiş mısırın potansiyel olumsuz etkileri tartışılmakta iken, mısırda insan sağlığı için gözden kaçmaması gereken en büyük tehdit mikotoksinlerdir. Bu çalışmada, Türkiye‘nin 24 ilinde bulunan 552 köyün 634 tarlasından toplanan mısır numuneleri GDO, aflatoksin B1, toplam aflatoksin, fumonisin B1, fumonisin B2, T-2 toksin, HT-2 toksin, zearalenon ve deoksinivalenol yönünden incelenmiştir. Tarama yapılan mısır numunelerinin hiçbirinde transgenik elemente rastlanmamıştır. Analiz edilen 634 numunenin sadece bir tanesinde Türk Gıda Kodeksi limitinin üzerinde aflatoksin B1 miktarına rastlanmış, numunelerin toplam aflatoksin değerleri Türk Gıda Kodeksi limitinin altında çıkmıştır. Paçal oluşturulan numunelerin hiçbirinde T-2 toksin, HT-2 toksin, zearalenon ve deoksinivalenol tespit edilmezken, fumonisin tespit edilen numunelerin toplam fumonisin miktarı Türk Gıda Kodeksi limitinin altında bulunmuştur. Bu sonuçlar, yerli üretim mısırların gıda güvenliği açısından uygun standartları sağladığını göstermektedir.

Mycotoxins are secondary metabolites produced by various fungi and are known to have a significant negative impact on human and animal health. When feedstuffs are contaminated with mycotoxins, their toxicities may be caused a variety of diseases. In this study, the residual mycotoxins in feedstuffs were analyzed using LC–MS/MS incorporated with QuEChERS extraction. Analytical method validation was performed for LOD, LOQ, linearity, and recoveries with consideration of matrix effects prior to the residual analysis. They were all reached to the accepted range of validation level. Using 39 feedstuff samples (5 g) for mycotoxin analysis, nine samples were contaminated by four major mycotoxins such as fumonisin B1 (FB1), deoxynivalenol, fumonisin B2, and zearalenone. Among them, FB1 was detected at the highest concentration as 18.0943 mg/kg. The total sum of fumonisins in 39 samples did not exceed the maximum residual level (MRL) criterion set by Korean Food and Drug Administration. Altogether, intensive management of mycotoxins in Korean feedstuffs should be implemented with proper and routine monitoring, even their residual concentrations are not exceeded over the MRL levels because of high frequent detection found in this study.

Fusarium verticillioides is a prolific producer of useful secondary metabolites such as naphthoquinone pigments, monoterpenes, and sesquiterpenes, as well as the harmful mycotoxins fumonisins. A strategy to increase their production includes creating a proper nutritional environment that enables the fungus to produce the compounds of interest. The aim of the present work was to study the effect of different carbon sources (glucose, fructose, xylose, sucrose, and lactose) on secondary metabolites biosynthesis in F. verticillioides submerged cultures. The production of volatile terpenes was evaluated through gas chromatography coupled to mass spectrometry. The quantification and identification of pigments was conducted using a UV/VIS spectrophotometer and NMR spectrometer, respectively. The quantification of fumonisin B1 and fumonisin B2 was performed by high-performance liquid chromatography. Our results showed that the biosynthesis of naphthoquinone pigments, monoterpenes, and sesquiterpenes was highest in cultures with fructose (13.00 ± 0.71 mmol/g), lactose [564.52 × 10−11 ± 11.50 × 10−11 μg/g dry weight (DW)], and xylose (54.41 × 10−11 ± 1.55 × 10−11 μg/g DW), respectively, with fumonisin being absent or present in trace amounts in the presence of these carbon sources. The highest biosynthesis of fumonisins occurred in sucrose-containing medium (fumonisin B1: 7.85 × 103 ± 0.25 × 103 μg/g DW and fumonisin B2: 0.38 × 103 ± 0.03 × 103 μg/g DW). These results are encouraging since we were able to enhance the production of useful fungal metabolites without co-production with harmful mycotoxins by controlling the carbon source provided in the culture medium.

Along with Aspergillus and Penicillium spp., mycotoxins produced by Fusarium species are important contaminants in food and feed industry. This study was designed to investigate the biodegradation potential of a collection of Bacillus spp. In different substrates artificially contaminated with single or multiple Fusarium mycotoxins. B. megaterium BM344-1 and B. pumilus BP344-3 showed the highest degradation (100%) of zearalenone (ZEN) on liquid Luria Bertani (LB) medium. Bacillus cereus BC344-2, Bacillus cereus BC399-2, Bacillus simplex BS350-3 and Paenibacillus favisporus PF341, showed significant biodegrading capacity on T-2 toxin, leading to 72, 70, 71 and 88% decontamination, respectively. Complete degradation (100%) of ZEN was observed at a contamination level of 0.375 μg/ml in LB, while showing a decreasing trend of degradation (87 and 84%) at increasing levels of mycotoxins (0.750 and 1.50 μg/ml LB medium, respectively). Likewise, BM344-1 showed a decreasing degradation trend (81, 78% and 55%) at increasing T-2 contamination levels (0.3, 0.6 and 1.2 μg/ml, respectively). The ability of BM344-1 to degrade fumonisin B1 (FB1) and fumonisin B2 (FB2) was 14 and 12%, respectively. In LB medium concurrently contaminated with multiple mycotoxins (ZEN + T-2+HT-2+FB1+FB2), the degradation ability of BM344-1 was not affected by low-to-medium levels of each mycotoxin, while it was significantly lowered upon addition of each toxin at higher levels. In vivo mycotoxin degradation potential of BM344-1 tested on artificially contaminated maize showed 25–44% reduction of ZEN and 15–19% reduction of T-2, with a non-significant effect in case of combined mycotoxins contamination. B. megaterium BM344-1, having significant biodegrading properties on multiple Fusarium mycotoxins, can be considered for its potential application as a mycotoxin decontamination agent in the food industry.

Edible insects are widely consumed in Africa, Asia, Oceania and Latin America, but less commonly so in Western countries. Since the turn of the millennium, however, entomophagy has aroused growing interest worldwide in response to the increasing scarcity of food resources. In fact, edible insects can be a source of high-quality protein, and also of fat, energy, minerals and vitamins. However, the lack of regulatory guidelines for microbiologically or chemically hazardous agents potentially present in these new foods (e.g., mycotoxins) may make their consumption unsafe. In this work, we developed an environmentally friendly analytical method using natural deep eutectic solvents (NADES or natural DES) in combination with ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of six mycotoxins of great concern owing to their toxic effects on humans and animals (namely, fumonisin B1, fumonisin B2, T-2 toxin, HT-2 toxin, ochratoxin A and mycophenolic acid) in insect-based food products. The target mycotoxins were co-extracted from cricket flour by using the optimum DES composition (namely, a mixture of choline chloride and urea, in a 1:2 mole ratio, containing 15% water which resulted in the highest extraction recoveries for all toxins). An experimental design method (Fractional Factorial Design (FFD) was used to examine the influence of the operational variables DES volume and water content, amount of sample, extraction time and extraction temperature on the extraction efficiency for each mycotoxin. Under optimum conditions, extraction recoveries were close to 100% except for fumonisin B2 (70%) and T-2 toxin (50%), with relative standard deviations (RSDs) below 13% in all cases. The proposed NADES-UHPLC–MS/MS method was validated in accordance with the European Commission 2002/657/EC and 2006/401/EC decisions, and used to determine the target compounds in cricket flour, silkworm pupae powder and black cricket powder., This work was funded by the Spanish Ministry of Economy and Competitiveness (MINECO, Projects RTI2018-096410-B-C21 and AGL2017-89417-R). F.P. acknowledges award of a pre-doctoral FPU grant by the Ministry of Education and Vocational Training, and G.A.-R. a “Juan de la Cierva” postdoctoral grant by MINECO.

Maize is an important worldwide commodity susceptible to fungal contamination in the field, at harvest, and during storage. This work aimed to determine the occurrence of Fusarium spp. in maize grains produced in the Tagus Valley region of Portugal and the levels of related mycotoxins in the 2018 harvest and during their storage for six months in barrels, mimicking silos conditions. Continuous monitoring of temperature, CO2, and relative humidity levels were done, as well as the concentration of mycotoxins were evaluated and correlated with the presence of Fusarium spp. F. verticillioides was identified as the predominant Fusarium species. Zearalenone, deoxynivalenol and toxin T2 were not found at harvest and after storage. Maize grains showed some variability in the levels of fumonisins (Fum B1 and Fum B2). At the harvest, fumonisin B1 ranged from 1297 to 2037 µg/kg, and fumonisin B2 ranged from 411 to 618 µg/kg. Fumonisins showed a tendency to increase (20 to 40%) during six months of storage. Although a correlation between the levels of fumonisins and the monitoring parameters was not established, CO2 levels may be used to predict fungal activity during storage. The composition of the fungal population during storage may predict the incidence of mycotoxins.

The prevalence of mycotoxins is often increased by the climatic conditions prevailing in tropical regions. Reports have revealed the contamination of mycotoxins in some types of vegetable oil. However, vegetable oil is one of the essential ingredients used in food preparation. Thus, this study determined the occurrence of multi-mycotoxins in six types of vegetable oils commercially available in Thailand to assess the consumer health risk. In total, 300 vegetable oil samples (olive oil, palm oil, soybean oil, corn oil, sunflower oil, and rice bran oil) collected from various markets in Thailand were analyzed for the presence of nine mycotoxins, namely, aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), beauvericin (BEA), ochratoxin A (OTA), zearalenone (ZEA), fumonisin B1 (FB1), and fumonisin B2 (FB2) using a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based procedure and a triple quadrupole mass spectrometer equipped with an electrospray ionization source. The incidences of mycotoxin contamination varied among the different types of oil samples. AFB1, AFB2, ZEA, FB1, and FB2 were most frequently found in contaminated samples. AFB2, BEA, ZEA, FB1, and FB2 contaminated olive oil samples, whereas AFB1, AFB2, AFG2, and OTA contaminated palm oil samples. AFB1, AFB2, and ZEA were found in soybean oils, whereas ZEA, FB1, and FB2 contaminated corn oil samples. AFB1 and AFG1 contaminated sunflower oil samples, whereas AFB1, AFB2, AFG1, and OTA were detected in rice bran oil samples. However, the contamination levels of the analyzed mycotoxins were below the regulatory limits.

In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of Aspergillus- and Penicillium-mycotoxins, (2) low levels of fumonisins and other Fusarium-mycotoxins, (3) high levels of Aspergillus-mycotoxins, (4) high levels of non-regulated Fusarium-mycotoxins, (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some Fusarium mycotoxins, together with the tetrapeptide tentoxin (an Alternaria toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese.

Mycotoxin-producing Aspergilli (Circumdati, Flavi, and Nigri), usually associated with contaminated food, may also cause respiratory disorders and are insufficiently studied in water-damaged indoor environments. Airborne (N = 71) and dust borne (N = 76) Aspergilli collected at post-flood and control locations in Croatia resulted in eleven different species based on their calmodulin marker: A. ochraceus, A. ostianus, A. pallidofulvus, A. sclerotiorum, and A. westerdijkiae (Circumdati), A. flavus (Flavi), and A. tubingensis, A. welwitschiae, A. niger, A. piperis, and A. uvarum (Nigri). Most of the airborne (73%) and dust borne (54%) isolates were found at post-flood locations, and the highest concentrations measured in indoor air (5720 colony-forming units (CFU)/m3) and dust (2.5 &times, 105 CFU/g) were up to twenty times higher than in the control locations. A. flavus dominated among airborne isolates (25%) at the unrepaired locations, while 56% of the dust borne Aspergilli were identified as A. tubingensis and A. welwitschiae. The ability of identified isolates to produce mycotoxins aflatoxin B1 (AFB1), fumonisin B2 (FB2), and ochratoxin A were assessed by LC-MS analysis. All ochratoxin A (OTA)-producing Circumdati belonged to A. westerdijkiae (13.7 &plusmn, 15.81 &micro, g/mL), in the section, FlaviA. flavus produced AFB1 (2.51 &plusmn, 5.31 &micro, g/mL), while A. welwitschiae and A. niger (section Nigri) produced FB2 (6.76 &plusmn, 13.51 &micro, g/mL and 11.24 &plusmn, 18.30 &micro, g/mL, respectively). Water damage dominantly supported the occurrence of aflatoxigenic A. flavus in indoor environments. Yet unresolved, the causal relationship of exposure to indoor Aspergilli and adverse health effects may support the significance of this research.

Coix seed is an important food and traditional Chinese medicine in China and other Asian countries. Notably, coix seed is currently being used as a traditional medicine for the treatment of COVID-19 in China. However, coix seeds are generally contaminated by mycotoxins, and this risk cannot be ignored. In this paper, we developed a method that involves direct extraction and UHPLC-HRMS analysis for the simultaneous detection of 24 mycotoxins in coix seeds. UHPLC-HRMS instrument and data acquisition parameters, and the sample pretreatment were optimised. One-step extraction showed several advantages compared to the three commercial solid-phase extraction clean-up methods, including ease of use, reduced time of sample preparation, low cost, good recovery, and acceptable matrix effect. The method validation results indicate that all mycotoxins have good linearity and sensitivity. Recoveries were between 74.2-101.1%, and RSD ranged from 0.1-5.8%. The LOQs for 24 mycotoxins were in the range of 0.5-100 µg/kg. To survey the contamination levels of these mycotoxins in commercial coix seeds, more than 70 samples were collected from Chinese markets and were analysed using the newly developed method. Zearalenone (positive ratio: 98.7%, range:1.1-1562 µg/kg), deoxynivalenol (positive ratio: 87%, range: 8.4-382.5 µg/kg), nivalenol (positive ratio: 85.7%, range: 26.8-828.2 µg/kg), fumonisin B1 (positive ratio: 84.4%, range:2.5-314.5 µg/kg), fumonisin B2 (positive ratio: 75.3%, range:1.6-72.8 µg/kg), fumonisin B3 (positive ratio: 48%, range:1.0-203.6 µg/kg), aflatoxin B1 (positive ratio: 29.9%, range: 0.39-14.7 µg/kg), sterigmatocystin (positive ratio: 29.9%, range: 1.4-51.6 µg/kg), and tenuazonic acid (positive ratio: 19.5%, range 36.1-105.7 µg/kg) were the most frequent mycotoxin contaminants. These results highlight the importance of routine monitoring and control of mycotoxins in coix seeds.

The occurrence of diseases in the cultivation of maize (Zea mays L.) leads to the loss of grain yield and the simultaneous deterioration of its quality. Fungi of the genus Fusarium spp. pose the greatest threat to maize cultivation. These fungi occur at all stages of the plant&rsquo, s development, causing Fusarium seedling blight, root rot, foot rot, and ear rot, including grains. Therefore, the aim of the conducted field studies was to determine the influence of selected agriculture factors on mycotoxin contents in maize grain and ear core. Moreover, it should be noted that the presence of mycotoxins in food products in terms of legal regulations is quite a fundamental barrier both in domestic and international trade. The field experiment was carried out at the Department of Agronomy of Poznań University of Life Sciences, on the fields of the Experimental and Educational Unit in Swadzim, Poland, in 2013&ndash, 2014, in four field replicates. The study involved the following factors: soil sowing preparation method, types of variates, and fertilization method, all of which shape mycotoxin accumulation. The results indicated that the main effects of years were significant for all observed traits for both the core and grain. The concentrations of deoxynivalenol, nivalenol, and zearalenone were higher in 2014 than in 2013 for both core and grain. The concentrations of fumonisin B1 and fumonisin B2 were higher in the first year of study for core and grain. The direct sowing of maize significantly affects plants&rsquo, health status, expressed by an increase in mycotoxin accumulation. Traditional hybrid SY Cooky characterized higher values of observed traits than &ldquo, stay-green&rdquo, hybrid Drim (except fumonisin B2 for cob). The positive effect of the row method of NP fertilizer application is based on a decrease in mycotoxin content.

Fumonisin contaminates food and feed extensively throughout the world, causing chronic and acute toxicity in human and animals. Currently, studies on the toxicology of fumonisins mainly focus on fumonisin B1 (FB1). Considering that FB1, fumonisin B2 (FB2) and fumonisin B3 (FB3) could coexist in food and feed, a study regarding a single toxin, FB1, may not completely reflect the toxicity of fumonisin. The gastrointestinal tract is usually exposed to these dietary toxins. In our study, the human gastric epithelial cell line (GES-1) was used as in vitro model to evaluate the toxicity of fumonisin. Firstly, we found that they could cause a decrease in cell viability, and increase in membrane leakage, cell death and the induction of expression of markers for endoplasmic reticulum (ER) stress. Their toxicity potency rank is FB1 &gt, FB2 &gt, &gt, FB3. The results also showed that the synergistic effect appeared in the combinations of FB1 + FB2 and FB1 + FB3. Nevertheless, the combinations of FB2 + FB3 and FB1 + FB2 + FB3 showed a synergistic effect at low concentration and an antagonistic effect at high concentration. We also found that myriocin (ISP-1) could alleviate the cytotoxicity induced by fumonisin in GES-1 cells. Finally, this study may help to determine or optimize the legal limits and risk assessment method of mycotoxins in food and feed and provide a potential method to block the fumonisin toxicity.

Climate change is primarily manifested by elevated temperature and carbon dioxide (CO2) levels and is projected to provide suitable cultivation grounds for pests and pathogens in the otherwise unsuitable regions. The impacts of climate change have been predicted in many parts of the world, which could threaten global food safety and food security. The aim of the present work was therefore to examine the interacting effects of water activity (aw) (0.92, 0.95, 0.98 aw), CO2 (400, 800, 1200 ppm) and temperature (30, 35 &deg, C and 30, 33 &deg, C for Fusarium verticillioides and F. graminearum, respectively) on fungal growth and mycotoxin production of acclimatised isolates of F. verticillioides and F. graminearum isolated from maize. To determine fungal growth, the colony diameters were measured on days 1, 3, 5, and 7. The mycotoxins produced were quantified using a quadrupole-time-of-flight mass spectrometer (QTOF-MS) combined with ultra-high-performance liquid chromatography (UHPLC) system. For F. verticillioides, the optimum conditions for growth of fumonisin B1 (FB1), and fumonisin B2 (FB2) were 30 &deg, C + 0.98 aw + 400 ppm CO2. These conditions were also optimum for F. graminearum growth, and zearalenone (ZEA) and deoxynivalenol (DON) production. Since 30 &deg, C and 400 ppm CO2 were the baseline treatments, it was hence concluded that the elevated temperature and CO2 levels tested did not seem to significantly impact fungal growth and mycotoxin production of acclimatised Fusarium isolates. To the best of our knowledge thus far, the present work described for the first time the effects of simulated climate change conditions on fungal growth and mycotoxin production of acclimatised isolates of F. verticillioides and F. graminearum.

An analytical method using immunoaffinty column and HPLC was developed for determination fumonisins in corn and corn products. The detection limits for fumonisin B1 (FB1) and fumonisin B2 (FB2) were 0.03 ppm and 0.07 ppm, respectively. Spiked FB1 in dried corn (kernel and flour), fresh corn, corn snack and corn flake are at 0.2-3.0 ppm level, with recovery ranged from 79.2-108.8% and relative standard deviation (RSD) of 2.9-21.9%. FB2 spiked under the same condition as FB1 had recovery ranges of 70.0-106.0% and RSD of 4.7-20.0%. A total of 76 samples were collected, including 20 dried corn (kernel or flour), 5 fresh corn, 15 corn snacks, 10 corn flakes, 5 corn starch, 5 canned corn from commercial markets and 16 raw corn materials were purchased from corn snack manufacturers. From the total 76 samples, 11 samples (14.5%) were detected with FB1 and/or FB2. Seven samples of corn were at 0.05-0.13 ppm level, 1 fresh corn with 0.15 ppm, 2 corn snacks with 0.5 and 0.16 ppm, respectively, where as 1 corn raw material was at the 0.09 ppm level. The highest contamination was only 0.16 ppm. Nevertheless, no fumonisin was detected in corn flake, corn starch and canned corn samples.

Fumonisins contamination of food commodities is a worldwide problem, especially for maize. The ability to produce fumonisinsis a trait of several species of Fusarium, mainly F. verticillioides and F. proliferatum on maize, and some Aspergillus species.A. nigerand its sister speciesA. welwitschiae, can contribute to fumonisin B2(FB2) accumulation in maize kernels, although to a lesser extent than fumonisin-producingFusariumspecies. Fumonisins risk monitoring represents an effective strategy in the integrated approach for mycotoxin risk management and reduction. The availability of a user-friendlymolecular assay for the detection oftoxigenic fungal species represents a valuable tool in understanding and managing upcoming mycotoxin contamination. In this study, we developed a LAMP assay, based on the detection offum10, for a rapid and specific molecular detection of FB2-producingA. nigerandA. welwistchiae, potentially useful to perform monitoring directly “on site” in maize chain. Results showed that very low amounts of conidia are suitable to detect the presence of the target gene, thus providing information about the presence of FB2-producingAspergillusspecies and the possible upcoming fumonisins contamination in maize. The assay was combined with a suitable protocol for “in field”crude DNA extraction and a colorimetric method for easy naked-eye evaluationof results, offering a reliable and user-friendly tool to support effective reduction strategies of mycotoxin contamination in crop management programs.

Investigating the in vitro fumonisin biosynthesis and the genetic structure of Fusarium verticillioides populations can provide important insights into the relationships between strains originating from various world regions. In this study, 90 F. verticillioides strains isolated from maize in five Mediterranean countries (Italy, Spain, Tunisia, Egypt and Iran) were analyzed to investigate their ability to in vitro biosynthesize fumonisin B1, fumonisin B2 and fumonisin B3 and to characterize their genetic profile. In general, 80% of the analyzed strains were able to biosynthesize fumonisins (range 0.03&ndash, 69.84 &mu, g/g). Populations from Italy, Spain, Tunisia and Iran showed a similar percentage of fumonisin producing strains (&gt, 90%), conversely, the Egyptian population showed a lower level of producing strains (46%). Significant differences in fumonisin biosynthesis were detected among strains isolated in the same country and among strains isolated from different countries. A portion of the divergent FUM1 gene and of intergenic regions FUM6-FUM7 and FUM7-FUM8 were sequenced to evaluate strain diversity among populations. A high level of genetic uniformity inside the populations analyzed was detected. Apparently, neither geographical origin nor fumonisin production ability were correlated to the genetic diversity of the strain set. However, four strains from Egypt differed from the remaining strains.

Fumonisins (FB1+FB2) and deoxynivalenol (DON) are mycotoxins produced by Fusarium species that might be present in maize and maize products. Knowledge on their occurrence in nixtamalized maize from Mexico together with an accompanying risk assessment are scarce, while nixtamalized maize is an important food in Mexico. This study presents the occurrence of FB1 + FB2 and DON in nixtamalized maize samples collected in Mexico City and analyses their distribution and resulting estimated daily intake for Mexican consumers by a probabilistic approach using a two-dimensional Monte-Carlo simulation. The results obtained reveal that for FB1 + FB2, 47% of the Mexican men and 30% of the Mexican women might exceed the provisional tolerable daily intake (PMTDI) of 2 &micro, g/kg bw/day for fumonisins and for DON, 9% of men and 5% of women would be exceeding the PMTDI of 1 &micro, g/kg bw/day, corresponding to the high consumers. The results raise a flag for risk managers in Mexico, to consider regulations and interventions that lower mycotoxin levels in nixtamalized maize for human consumption.

In our study, a reliable and rapid analytical method for the simultaneous determination of 15 mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, alternariol, agroclavine, citrinin, diacetoxyscirpenol, deoxynivalenol, fumonisin B1, fumonisin B2, ochratoxin A, sterigmatocystin, T-2 toxin, and zearalenone) in liquorice using ultra-HPLC coupled to tandem MS was developed and validated. Due to the complex ingredients in liquorice, we chose a QuEChERS-based extraction procedure as the sample pretreatment. Meanwhile, for the first time, acetate buffer was used to replaced water, which can greatly reduce the concentration of formic acid in acetonitrile, which further reduces the extraction efficiency of impurities. The optimal combination of adsorbents is 150 mg primary secondary amine, 150 mg silica gel, 600 mg octadecylsilane, and 900 mg anhydrous magnesium sulfate. Electrospray ionization in both positive- and negative-ionization modes was applied to detect all the mycotoxins in a single run time of 15 min, with LOQs in the range of 0.125–2.5 μg/kg. The recoveries of determination obtained were in the range of 81.0–104.7%, whereas the analytes could be accurately quantified in the 0.25–625 μg/kg concentration range, with all coefficients being >0.992. Intra- and interday reproducibility were lower than 5.5 and 8.9%, respectively, for all analytical mycotoxins. The validated method was finally applied to screen mycotoxins in 31 batches of real samples collected from drugstores and hospitals in Shanghai, China. Our survey findings show that six mycotoxins were detected, including alternariol, citrinin, deoxynivalenol, fumonisin B1, ochratoxin, and zearalenone, and that the positive rate of mycotoxins was 54.8% in real samples, ranging from 3.37 to 520.6 μg/kg.